108 BIOLOGICAL OCEANOGRAPHIC PROCESSES

in which ^X(T,I) and KS(T) indicate the maximum specific growth rate controlled by
temperature and light intensity, and the half-saturation constant controlled by temperature,
respectively. For the possible relation between the maximum specific growth rate and
temperature, the Arrhenius equation was evaluated by using a set of literature values on
continuous culture experiments with freshwater and marine phytoplankton, and found to fit
nicely. Equation obtained was , = A c-T (75)
where A is constant (d~l), is the activation energy (cal-mole-1), R is the universal gas constant
(cal-K-1 mole"1) and Tis temperature in Kelvin scale (K). Light intensity gives a strong effect
on the activation energy, E, then shown by E(I) representing lightdependent activation energy.
However, actual relations cannot be formulated due to incomplete data available at this stage.
This is also the case for the temperature effect on Ks; KS(T).

Once functional relations between two variables, Mmax and Ks, and the environmental
parameters, nutrient concentrations, temperature and light intensity, are known for each
phytoplankton species, the specific growth rate for each species, 9 can be estimated at a given
condition controlled by the environmental parameters mentioned above. The actual growth of a
given phytoplankton species population can then be estimated by the application of the growth
rate obtained into a growth equation such as eqn. (55). With the aid of a computer, growth
calculation can be made with minimum efforts even though actual calculation is highly
complicated. However, care must be taken to overgeneralize and indiscriminately apply models,
because computer calculations often go further than they should.

3.1.7 Chemosynthesis
Some micro-organisms can satisfy their primary energy requirements by utilizing simple
inorganic compounds, such as ammonia, methane, or nitrite, or elements, such as ferrous iron,
hydrogen gas, or water-insoluble amorphous sulfur. All of the known organisms which comprise
this (chemosynthetic) group are bacteria. Most of their inorganic substrates are derived from the
decay of organic matter which is itself primarily formed through photosynthesis. Consequently,
if one follows the origin of the energy used in chemosynthesis, chemosynthetic processes may
not be considered as primary production. However, chemosynthesis usually involves carbon
dioxide fixation and the primary formation of new particulate material. Thus chemosynthesis
may be considered as a special case of primary production on the grounds of its trophic position
in the marine food chain.

In the past it was believed that the inorganic substrates reacted directly with molecular
oxygen to form oxidized end-products. However, Bunker (1936) showed that molecular oxygen
was not directly involved in sulfur oxidation by thiobacilli. The fact that oxygen in the end-
products originated from water resulted in a new concept of chemosynthesis, although in some
cases it is still questionable whether the concept is adaptable to all cases of chemosynthetic
activity. Thus reactions of the type suggested by Bunker involve dehydrogenations rather than
oxidations. In general terms the chemosynthetic process can be expressed conveniently in three
stages according to Bunker's concept (Gundersen, 1968).

1. As the result of dehydrogenation, high reducing power is produced as follows: 2 +

nH20dehydI0gcn r AO + 4n[H++ e r ] (76) (Inorganic substrates) (reducing power)
(oxidized end-product) where the symbol [H+ + e"] is used as a synonym for the reducing
power.
2. A proportion of the reducing power is then utilized for energy production (adenosine
triphosphate, ATP, synthesis) by being transferred through the cytochrome system to
molecular oxygen. A second part of the reducing power is transferred to NAD
(nicotinamide adenine dinucleotide) in order to produce reduced NAD (or NADH2,
reduced nicotinamide adenine dinucleotide). These relationships might be visualized as
follows: 4[H+ + e ] + wADP + m ?, + 0 2 cytochrome , _ A (7T7O)
3. system I 2 2[H+ + e"] + NAD 2H20 + Watp NADH2 (78) where ADP is adenosine
diphosphate, and Pj is inorganic phosphate. Some anaerobes can use bound oxygen
derived from inorganic compounds instead of free oxygen as shown in the above equation
(e.g. sulphate-reducing bacteria).

The ATP and NADH2 are then used for the assimilation of carbon dioxide:
12NADH2 + 18ATP + 6C02 - i C 6H 1 20 6 + 6H20 + 18ADP + 18Pi + 12NAD. (79) Thus the
different inorganic substrates used by chemosynthetic bacteria are not merely the sole source of
the organism's energy but also the sole source of their reducing power.

Depending on differences in the organic substrate (AH2), chemosynthetic bacteria are

classified into several groups, such as nitrifying, sulfur, hydrogen, methane, iron and carbon
monoxide bacteria. Table 23 shows some representative chemosynthetic bacteria inhabiting
marine environments. The overall efficiencies of chemosynthesis during the growth of the
bacteria (e.g. nitrifying or sulfur bacteria) can be expressed as the ratio between the total energy
consumed in carbon dioxide assimilation and the energy liberated by the primary inorganic
compounds during oxidation; these efficiencies are 6 to 8% (Gibbs and Schiff, 1960) although
the efficiency may sometimes change drastically with different stages of cultures. For example,
young cultures of Nitrosomonas gave values approaching 50%; in older cultures, the efficiency
dropped to 7% (Hofmann and Lees, 1952). Most chemosynthetic bacteria require free oxygen as
the electron acceptor in the second step described above. However, facultative or obligate
anaerobic bacteria, such as Thiobacillus denitrificand Desulfovibrio desulfricans, can use bound
oxygen derived from nitrate or sulfate.

Among inorganic substrates available for the chemosynthetic bacteria, nitrogen and sulfur
compounds are relatively abundant and widely distributed compared with the other reduced
compounds in the pelagic environment of the oceans. Among reduced nitrogen compounds,
ammonia may be present in concentrations up to ca. 5 Mg at/1 and nitrite at concentrations up to
ca. 2 g at/1 (Riley and Chester, 1971). Actual occurrences of nitrifying bacteria, expressed in
colonies per litre of sea water, were found to be < 1 in the north Atlantic Ocean, < 10 in the
Pacific Ocean, ca. 104 in the Indian Ocean near an island, and ca. 106 in Barbados harbor
(Watson, 1965; Hattori and Wada, 1971). Some of the strains of nitrifying bacteria isolated from
marine waters are the same or similar to those from fresh water or soil, but others are peculiar to
marine environments (e.g. see Watson, 1971). Nitrification by marine bacteria, using either
ammonia or nitrite as a substrate, is reported to be more efficient at low (<0 1 ml/1) oxygen
concentrations (Carlucci and McNally, 1969).
 The qualities of reduced sulfur compounds in the marine environments are generally
much greater than the quantities of reduced nitrogen compounds. Thiosulfate and polythionates
may sometimes be present at levels from 0 to 100 Mg at/1, the highest concentrations of these
compounds being detected near to shore (Tilton, 1968). Sulphides are generally not detected
within the analytical limit of < 1 Mg at/1 in open ocean waters (Tilton, 1968). The number of
colonies of sulfur bacteria (Thiobacillus spp.) have been found to range from 0 to 275 per 100 ml
(Tilton et al., 1967). These values for colony counts are several orders of magnitude smaller than
those (103 to 104 per 100 ml) which were calculated by Tilton et al. (1962) on the basis of the
amount of reduced sulfur compounds.

The reduced inorganic substrates are mainly produced through anaerobic metabolic
processes. Consequently, anaerobic environments, which sometimes develop in fjords and
estuaries, create favourable habitats for chemosynthetic bacteria. Under such conditions vigorous
growth of certain species of chemosynthetic bacteria (usually sulfur, hydrogen, or methane
bacteria) has been observed
TABLE 2 3 . SUMMARY OF CHEMOSYNTHETIC BACTERIA GROWING IN THE
OCEAN AND THEIR INORGANIC SUBSTRATES
Oxidized
Inorganic
substrate
endproduct
Oxidizer
AF*
(K cal)
Ability to grow
heterotrophically Habitat
(1) Nitrifying bacteria
Nitrobacter spp. N 0 2 N03 o 2 18 Soil, fresh, and sea waters
Nitrococcus mobilis WATSON & WATERBURG N 0 2 N 0 3 o 2 18 Sea water
Nitrospina gracilis WATSON & WATER BURG N 0 2 N 0 3 o 2 18 Sea water
Nitrosomonas spp. NH3 N 0 2 o 2 85 Soil, fresh, and sea waters
Nitrosococcus oceanus (WATSON) comb. nov. H3 or NH2OH N 0 2 o 2 85 Sea water
(2) Sulfur bacteria
Thi ovulum ma ja s HINZE H2S S 50 ? Sea water
Beggiatoa spp. H2S S 50 + Fresh and sea waters
Thiospira bipunctata MOLISCH H2S S 50 7 Sea water
Thiothrix spp. S s o 4- o 2 119 ? Soil, fresh, and sea waters
Thiobacillus thioparus BEIJERINCK 5/4 S2Ol 3/2 S 0 4- - + s o 2 112 + Soil, fresh, and sea
waters
Thiobacillus denitrificans BEIJERINCK 5S 5SO;- o 2 112 + Soil, fresh, and sea waters
(3) Hydrogen bacteria
Hydrogenomonas spp. H2 H20 o 2 56 + Soil, fresh, and sea waters
Desulfovibrio desulfricans (BEIJERINCK) H2 H20 s o 4 56 + Soil, fresh, and sea waters
KLUYVER & VAN NIEL
(4) Methane bacteria
Methanomonas spp. CH4 c o 2 o 2 + Soil, fresh, and sea waters
(5) Iron bacteria
Grallionella spp. 4FeC03 4Fe(OH)3 o 2 81 +
(6) Carbon monoxide bacteria
Sarcina bakerii SCHNELLEN CO CH4 H2 + Soil, fresh, and sea waters
*Free energies per number of moles of electron donor indicated. Values reviewed by Gibbs
and Schiff (1960) are mainly quoted here.

(Kuznetsov, 1959; Sokolova and Karavaiko,

1964).

It should be mentioned that an obligate anaerobic strain, Desulfovibrio desulfricans, has

been isolated even from oxic ocean waters (Kimata et al., 1955; Wood, 1958). In order to explain
this phenomenon, Baas Becking and Wood (1955)
proposed the idea of 'metabiosis' in which they suggested that several kinds of micro-organisms
coexist on or in bacterial aggregates and conditions for the production of reduced substrates
could occur within the aggregates. Thus the obligate anaerobic bacteria existing in oxic
environments are probably present within the microcosm of a
bacterial aggregate (Seki, 1972).

In the ocean, chemosynthetic activity can be estimated from carbon dioxide uptake in the
dark using NaH1 4C03 . Compared with carbon fixation through photosynthesis, dark C02
uptake is usually small (i.e. less than 5% of the photosynthesis on a daily basis within the
euphotic zone in pelagic areas). Dark C02 uptake is not entirely carried out by chemosynthesis,
but heterotrophic processes by
bacteria and algae (e.g. Wood and Werkman, 1935, 1936, and 1940) may result in the uptake of
small amounts of C 0 2 in the absence of light. Algal dependency on dark fixation of C 0 2 is low
(i.e. 3 to 5% of the total C02 fixed in the light) and this is about the same proportion of C 0 2
fixed by aerobic heterotrophic bacteria when they are growing on an organic substrate (Sorokin,
1961 and 1966). However, among facultative autotrophic bacteria which belong to an
intermediate metabolic type between obligate heterotrophs and chemoautotrophs, there is a
requirement of between 20 and 90% C02 during the oxidation of low molecular weight organic
compounds, such as methane and formic acid; for obligate chemoautotrophs the requirement for
C02 is close to 100%. Since in natural environments, micro-organisms which depend on different
kinds of substrates exist together, it is practically impossible to separate out and estimate the
actual activity of chemosynthetic organisms except at the time of vigorous growth of any one
species. However, a measure of dark C02 up take is a useful measure of chemosynthetic activity
in any environment. As an example, Seki(1968) studied seasonal changes in dark uptake of C02
in a small bay and showed that dark uptake of C02 near the bottom of the bay could be as high as
20% of the photosynthesis throughout the year and that during the spring dark uptake for Jhe
water column was sometimes 50% greater than photosynthesis.

Active chemosynthesis occurs in waters and sediments in which both aerobic and
anaerobic environments exist in the same column. Sorokin (1964a and b) has studied the
importance of chemosynthetic bacteria in the Black Sea, where a thick anaerobic zone exists
below the depths of ca. 150 m in the central part of the sea. A summary of his finding is shown in
Fig. 50. At depths shallower than 50 m, inorganic carbon is fixed through photosynthesis by
algae, and the daily photosynthesis
is reported to be ca. 350 mg C/m2/day during October. At the transition zone between 100 and
250 m, where environmental conditions are changing from aerobic to anaerobic, chemosynthetic
fixation of inorganic carbon is predominant. An in situ maximum value for daily chemosynthesis
was ca. 9mg C/m3/day; chemosynthesis for the water column amounted to approximately 200
mg C/m2/day. This active chemosynthesis is carried out mainly by sulfur bacteria (aerobic and
anaerobic Thiobacillus), and it is supported by a continual supply of reduced sulfur compounds,
such as H2S, S, and S203 , from the anaerobic zone. The potential activity of thiobacilli in water
samples taken from the depth of their maximum activity is very high compared with the in situ
activity; by determining the oxidation rate of thiosulfate added to a water sample, a potential
activity of 200 mg C/m3/day has been obtained.

Although the studies described above may appear to have limited geographical
significance, it is probable in fact that similar microzonation in environmental factors can occur
in many marine environments, especially in bays and estuaries. Also in all nearshore sediments
where wave action does not disturb the benthos, a high chemosynthetic activity will occur in a
depth zone of a few cm just below the sediment surface. If the transient zone of aerobic to
anaerobic conditions comes up above the depth receiving a few per cent of the surface
112 BIOLOGICAL OCEANOGRAPHIC PROCESSES
N . N 0 2 mg/m3 PHOTOSYNTHESIS mgC/m3/day
0 05 10 15 0 4 12 16
FIG. 5 0 . Daily chemosynthetic production and some factors influencing it in the Black Sea
(after Sorokin, 1964c).

illumination, anaerobic photosynthetic sulfur bacteria, which requires reductants (e.g. H2S, S)
for the hydrogen donor, grow vigorously. These organisms impart a purple or deep green colour
to the sediment or water column in which they exist.

3.2 HETEROTROPHIC PROCESSES

3.2.1 The Origin of Organic Substrates Heterotrophic organisms are dependent on an organic
carbon source to provide energy for growth. In the sea, most of the organic carbon utilized by
heterotrophic organisms originates from the marine biota, and only in near-shore coastal areas is
there any appreciable contribution of organic materials from the land. Exceptions to this
generalization are found where major rivers may influence the oceanic environment for a
considerable distance off shore; in particular, the Amazon River, with an annual discharge of 6 X
1012 m3 (or ca. 20% of the entire world wide river runoff), may contain sufficient organic
carbon (2 to 10 mg/1) to influence heterotrophic activity over an oceanic area approximately 107
km2 during
maximum runoff (Williams, 1968). Duursma(1965) recognized four main groups of organic
compounds which occur as dissolved substances in sea water. These are (1) nitrogen-free organic
matter, including carbohydrates, (2) nitrogenous substances,
including amino acids and peptides, (3) fatlike substances, and (4) complex substances, including
humic acids, derived from groups (1) and (2). In addition to these dissolved materials, particulate
organic materials also serve as a substrate for heterotrophic organisms. The nature and chemical
composition of these substances are
SIKLUS

ALLOCHTHONOUS ORGANIC MATERIALS

FROM THE LAND
c/
RESERVOIR OF AUTOCHTHONOUS
DISSOLVED AND PARTICULATE
ORGANIC MATERIALS IN THE SEA
Heterotrophic
uptake
I FILTER FEEDERS
I
-Excretion-
Heterotrophic
uptake
CARNIVORES - E x c r e t i o n -
BACTERIA
AND OTHER
HETEROTPCPHS
CO2 + INORGANIC
NUTRIENTS
.Hydrolysis
and autolysis

FIG. 51. Pathways of transfer and regeneration of organic

substrates in an aquatic ecosystem (modified from Johannes,
1968).

discussed in Section 1.4; their origin is illustrated in Fig. 51, which is adapted from Johannes
(1968). The figure illustrates that the two principal pathways of organic materials which act as
substrates for heterotrophic organisms result from the food chain of the sea, firstly through the
physiological release of materials and secondly
through the decomposition of plants and animals themselves. Autochthonous organic materials
derived by physiological processes include the release of dissolved organic materials by
phytoplankton; this subject has been reviewed by Fog